Neuregulin-4 attenuates diabetic cardiomyopathy by regulating autophagy via the AMPK/mTOR signalling pathway.

BACKGROUND: Diabetic cardiomyopathy is characterized by left ventricle dysfunction, cardiomyocyte apoptosis, and interstitial fibrosis and is a serious complication of diabetes mellitus (DM). Autophagy is a mechanism that is essential for maintaining normal heart morphology and function, and its dysregulation can produce pathological effects on diabetic hearts. Neuregulin-4 (Nrg4) is an adipokine that exerts protective effects against metabolic disorders and insulin resistance. The aim of this study was to explore whether Nrg4 could ameliorate DM-induced myocardial injury by regulating autophagy. METHODS: Four weeks after the establishment of a model of type 1 diabetes in mice, the mice received Nrg4 treatment (with or without an autophagy inhibitor) for another 4 weeks. The cardiac functions, histological structures and cardiomyocyte apoptosis were investigated. Autophagy-related protein levels along with related signalling pathways that regulate autophagy were evaluated. In addition, the effects of Nrg4 on autophagy were also determined in cultured primary cardiomyocytes. RESULTS: Nrg4 alleviated myocardial injury both in vivo and in vitro. The autophagy level was decreased in type 1 diabetic mice, and Nrg4 intervention reactivated autophagy. Furthermore, Nrg4 intervention was found to activate autophagy via the AMPK/mTOR signalling pathway. Moreover, when autophagy was suppressed or the AMPK/mTOR pathway was inhibited, the beneficial effects of Nrg4 were diminished. CONCLUSION: Nrg4 intervention attenuated diabetic cardiomyopathy by promoting autophagy in type 1 diabetic mice. Additionally, Nrg4 induced autophagy via the AMPK/mTOR signalling pathway.

Mycoplasma gallisepticum (HS strain) surface lipoprotein pMGA interacts with host apolipoprotein A-I during infection in chicken.

The adhesin protein from Mycoplasma gallisepticum (HS strain), namely pMGA1.2, is required for M. gallisepticum (MG) infection in chicken. However, the host factor(s) that interact with pMGA1.2 is not known. In this study, we prepared the membrane fraction of trachea epithelial cells from chicken embryos. Using an improved virus overlay protein blot assay (VOPBA) and glutathione S-transferase (GST) pull-down assay, we found that pMGA1.2 specifically bound to a ∼30 kDa host protein. This host protein was further identified by mass spectrometry as chicken apolipoprotein A-I (ApoA-I). We expressed and purified the recombinant ApoA-I protein in Escherichia coli and confirmed that it bound to the purified pMGA1.2 protein in vitro. Transiently expressed pMGA1.2 and ApoA-I were colocalized in HeLa cells. Finally, we designed small interfering RNA (siRNA) molecules to knock down the expression of either ApoA-I or pMGA1.2, which inhibited the MG-induced cell cycle disruption in cells of chicken embryo fibroblast cell line (DF-1). Similarly, knockdown of ApoA-I inhibited the cilia loss and damage in chicken trachea cells in MG infection. In summary, ApoA-I may be an essential host factor in MG infection through interacting with pMGA1.2.

Extracellular HMGB1 as Inflammatory Mediator in the Progression of Mycoplasma Gallisepticum Infection.

High-mobility group box 1 (HMGB1), a member of damage-associated molecular patterns (DAMPs), is involved in the immune regulation of several infectious diseases. Mycoplasma gallisepticum (MG) infection is proved to cause an abnormal immune response, but the role of HMGB1 in MG-induced chronic respiratory disease (CRD) is unclear. In this study, we found that HMGB1 was released from the nucleus to the extracellular in macrophages upon infection with MG. Extracellular HMGB1 bound to TLR2 activating the NF-κB pathway triggering a severe inflammatory storm and promoting the progression of MG infection. More importantly, TLR4 could be activated by HMGB1 to trigger immune disorders after TLR2 was silenced. This disease process could be interrupted by ethyl pyruvate (EP) inhibition of HMGB1 release or glycyrrhizic acid (GA). Furthermore, treatment of MG-infected chickens with GA significantly alleviated immune organ damage. In conclusion, we demonstrate that HMGB1 is secreted extracellularly to form an inflammatory environment upon MG infection, triggering a further cellular inflammatory storm in a positive feedback approach. Blocking MG-induced HMGB1 release or suppression downstream of the HMGB1-TLR2/TLR4 axis may be a promising novel strategy for the treatment of CRD. Furthermore, this study may provide a theoretical reference for understanding non-LPS-activated TLR4 events.

Evaluation of Glycyrrhizic Acid Therapeutic Effect and Safety in Mycoplasma gallisepticum (HS Strain)-Infected Arbor Acres Broilers.

This study was conducted to evaluate the therapeutic effects and safety of GA in MG-infected broilers. Our results showed that the minimum inhibitory concentration of GA was 31.25 µg/mL. Moreover, GA inhibited the expression of MG adhesion protein (pMGA1.2) in the broilers' lungs. GA treatment clearly decreased the morbidity of CRD and mortality in the MG-infected broilers. Compared with the model group, GA treatment significantly decreased gross air sac lesion scores and increased average weight gain and feed conversion rate in the MG-infected broilers. Histopathological examination showed GA treatment attenuated MG-induced trachea, immune organ and liver damage in the broilers. Moreover, GA treatment alone did not induce abnormal morphological changes in these organs in the healthy broilers. Compared with the model group, serum biochemical results showed GA treatment significantly decreased the content of total protein, albumin, globulin, alanine aminotransferase, aspartate aminotransferase, total bilirubin, creatinine, uric acid, total cholesterol, and increased the content of albumin/globulin, alkaline phosphatase, apolipoprotein B and apolipoprotein A-I. In conclusion, GA displayed a significant therapeutic efficacy regarding MG infection and had no adverse effects on the broilers (100 mg/kg/d).

High-normal thyroid function and risk of recurrence of atrial fibrillation after catheter ablation.

BACKGROUND: It has been shown that the concentration of serum free thyroxine (FT(4)) is independently associated with atrial fibrillation (AF), even in euthyroid persons. This study investigated the effect of a high-normal level of FT(4) on recurrence after catheter ablation of AF. METHODS AND RESULTS: The 244 consecutive patients with paroxysmal AF and who underwent circumferential pulmonary vein isolation (PVI) were prospectively enrolled. Exclusion criteria included prior or current thyroid dysfunction on admission, amiodarone medication for 3 months before admission. After a mean follow-up of 416+/-204 (91-856) days, the recurrence rates were 14.8%, 23.0%, 33.3%, 38.7% from the lowest FT(4) quartile to the highest FT(4) quartile, respectively (P=0.016). After adjustment for age, sex, left atrial diameter, and PVI, there was an increased risk of recurrence in the subjects with the highest FT(4) quartile compared with those with the lowest quartile (hazard ratio 3.31, 95% confidence interval 1.45-7.54, P=0.004). As a continuous variable, FT(4) was also an independent predictor of recurrence (hazard ratio 1.10, 95% confidence interval 1.02-1.18, P=0.016). CONCLUSIONS: Patients with high-normal thyroid function were at an increased risk of AF recurrence after catheter ablation.

Curing piglets from diarrhea and preparation of a healthy microbiome with Bacillus treatment for industrial animal breeding.

High-throughput farming of animals for an essential purpose such as large scale health and production of hogs is a challenge for the food industry in the modern world. The problem is that the breeding of livestock for fast growth or high yields of meat is often associated with illness and microbial infection that develop under the breeding conditions. Piglet diarrhea is most common pig disease, leading to heavy mortality and thereby economic loss. We proved that chemical drugs can relieve the symptoms of diarrhea in ill piglets, but they do not treat the underlying cause, i.e. significantly altered bacterial gut flora. Using Illumina sequencing of fecal DNA, we showed that the bacterial gut flora of piglets treated with antibiotics remain close to the ill conditions. However, using Illumina sequencing of fecal DNA from piglets treated with a specific Bacillus (Bacillus subtilis Y-15, B. amyloliquefaciens DN6502 and B. licheniformis SDZD02) demonstrated the efficiency of natural bioproducts not only on curing diarrhea, but also on beneficial bacteria to re-establish in the piglet gut. We therefore propose a new natural "medicine" to be explored by the world farm animal agriculture industry, particularly for sustainable improvement of swine livestock production and health.

Danon disease: Two patients with atrial fibrillation in a single family and review of the literature.

The present study reports on a family with two members affected by Danon disease but having different phenotypes. The clinical manifestations of Danon disease include cardiomyopathy, skeletal myopathy and different degrees of intellectual disability that varies greatly among patients. The present case study reports on two siblings, an older sister and a younger brother, with Danon disease from an affected pedigree, presenting with distinctly different phenotypes. The sister was diagnosed with dilated cardiomyopathy at the age of 26 years with an unfavorable outcome, while her younger brother presented with hypertrophic cardiomyopathy in a relatively stable state. The two probands shared the same mutation, c.974delTinsAA in exon 8, in the lysosomal-associated membrane protein-2 gene. Of note, the two patients had a pre-excitation pattern in the electrocardiogram on initial presentation and later developed atrial fibrillation (AF), which markedly aggravated heart failure. To the best of our knowledge, AF has not been widely reported in patients with Danon disease. The development of AF may have a prognostic value under these circumstances.

Can atrial vagal denervation influence ventricular function in a failing heart?

Atrial fibrillation (AF) and congestive heart failure (CHF) often coexist (AF-CHF), and each adversely affects the other with respect to management and prognosis. Therapy with antiarrhythmic drugs to maintain sinus rhythm was disappointing. Ablation is more successful than antiarrhythmic drug therapy for the prevention of AF with few complications, although in patients with AF-CHF it is noted. Ablating autonomic nerves and ganglia on the large vessels and the heart can result in AF suppression with little damage to healthy myocardium. Our study in patients with AF-CHF found that cardiac function aggravation was more frequent in patients with AF recurrence than that of those who successfully maintain sinus rhythm. The autonomic nervous system is a fine network spreading throughout the myocytes; hence the elimination of atrial vagal with radiofrequency catheter ablation can influence the innervation in sinus and AV nodes even in the ventricular region. Thus we propose that atrial vagal denervation may result in paratherapeutic sympathovagal imbalance in the ventricular region, which has a negative effect in a failing heart, although it is neutralized by the benefit accrued from sinus rhythm after successful ablation.

Can pulmonary vein antrum be defined by electrophysiological mapping?

Atrial fibrillation (AF) is the most common sustained arrhythmia, and treatments with anti-arrhythmia drugs (AADs) have been frustrating. Limitations of AADs prompted the development of percutaneous catheter ablation. In contrast to AADs, percutaneous catheter ablation offers the possibility of a lasting cure. The successful cure of AF by percutaneous catheter ablation comes from a widespread recognize that pulmonary vein antrum (PVA) plays an important role in the genesis and maintenance of AF, and circular ablation along the PVA can eliminate majority of AF. PVA is comprised of pulmonary vein-left atrium junctions. However, during ablation procedure, definition of PVA solely depends on angiography, and it is largely experience-dependent and there is a great deal of variation involved. Our study in patients with AF found that a unique potential with double deflections could be documented along PVA, but it cannot be recorded at PV side or LA side. Thus, we propose that documentation of PVA potentials can be used as a landmark to define PVA. Unlike angiography, documentation of PVA potentials can be objectively carried out by different operators, and the variations due to experience can be avoided.

[Asymptomatic recurrence of atrial tachy-arrhythmia after circumferential pulmonary vein isolation in patients with atrial fibrillation].

OBJECTIVE: To investigate the incidence, type, and predictors of asymptomatic relapse of atrial tachy-arrhythmia (ATa) after circumferential pulmonary vein isolation (CPVI) in patients with atrial fibrillation (AF). METHODS: Forty-eight consecutive patients with AF underwent CPVI and were followed up. Forty-eight hours Holter recording was performed 1, 3, and 6 months respectively after the initial CPVI procedure. Predictors of asymptomatic ATa relapse were determined by Logistic regression analysis for eight variables as follows: age, gender, AF type, existence of organic heart disease, diameter of left atria, left ventricular ejection fraction, procedure time, and heart rate variability after the procedure. RESULTS: Complete Holter data were acquired in 42 patients, 26 males and 16 females, aged: 58 +/- 14, including 25 patients with paroxysmal AF and 17 with non-paroxysmal AF. The standard deviations of R-R interval (SDNN) of the non-paroxysmal AF group was 92 ms +/- 19 ms, significantly longer than that of the paroxysmal AF group (78 ms +/- 15 ms, P = 0.011). The incidence of asymptomatic ATa recurrence rates 1, 3 and 6 months after CPVI were 8%, 12%, and 8% respectively in paroxysmal AF group and 23.5%, 29.4%, and 35.3% respectively in the non-paroxysmal AF group. The incidence of asymptomatic ATa recurrence 6 months after CPVI in the non-paroxysmal AF group was significant higher than that in the paroxysmal AF group (P < 0.05). AF was the dominant arrhythmia among the asymptomatic recurrence ATa, while atrial tachycardia constituted the major arrhythmia of the symptomatic recurrent ATa. CONCLUSION: (1) Asymptomatic ATa relapse is common among the patients undergoing CPVI. (2) The dominant type of asymptomatic recurrent arrhythmia is AF. (3) The independent predictors for asymptomatic ATa recurrence include non-paroxysmal AF, left atrial enlargement, and increase of SDNN.

Achievement of pulmonary vein isolation in patients undergoing circumferential pulmonary vein ablation: a randomized comparison between two different isolation approaches.

INTRODUCTION: Circumferential pulmonary vein ablation (CPVA) with the endpoint of pulmonary vein (PV) isolation has been developed as an effective therapy for atrial fibrillation (AF). This endpoint can be achieved either by closing gaps along circular lines or by segmental PV isolation inside the circular lines after creation of initial CPVA lesions. We investigated whether the clinical outcome depends on the PV isolation approach used during the first-time CPVA procedure. METHODS AND RESULTS: One hundred consecutive patients (69 male; age, 56.7 +/- 11.6 years) who underwent first-time CPVA for treatment of symptomatic AF were enrolled. PV isolation was randomly achieved either by CPVA alone (aggressive CPVA [A-CPVA] group, n = 50) or by a combination of CPVA with segmental PV ostia ablation (modified CPVA [M-CPVA] group, n = 50). Recurrence of atrial tachyarrhythmias (ATa) within 3 months after the initial procedure occurred in 30 patients (60%) in the M-CPVA group and in only 15 patients (30%) in the A-CPVA group (P < 0.01). ATa relapse after the first 3 months was detected in 21 patients (42%) in the M-CPVA group, compared with 9 patients (18%) in the A-CPVA group (P = 0.01). At 13 +/- 4 months, patients treated by the A-CPVA approach had greater freedom from ATa recurrence than patients who underwent M-CPVA (P = 0.01). The M-CPVA approach was the only independent predictor associated with procedural failure (RR 0.318; 95% CI 0.123-0.821; P = 0.02). CONCLUSIONS: When PV isolation is the endpoint of CPVA, the efficacy of the A-CPVA approach is better than that of M-CPVA.

Pulmonary vein tachycardia after pulmonary vein isolation in patients with atrial fibrillation.

BACKGROUND: Pulmonary vein (PV) isolation has been developed to treat patients with atrial fibrillation (AF), and the electrophysiological endpoint of PV isolation is the disappearance or dissociation of pulmonary vein potentials (PVPs). Pulmonary vein tachycardia (PVT) is the dissociated PV rhythm with a rapid rate. However, the characteristics and significance of PVT after pulmonary vein isolation in patients with AF remains unclear. METHODS: From June 2003 to June 2005, a total of 285 consecutive patients with drug refractory AF were included in this study, and they underwent segmental pulmonary vein ablation (SPVA) or circumferential pulmonary vein ablation (CPVA). PV isolation was the initial endpoint for both approaches with documenting disappearance or dissociation of PVPs. PVT was characterized as dissociated activities within PVs with a circle length (CL) of < 300 ms, and was classified into organized PVT or disorganized PVT according to the variance of CL. Systematic follow-up was conducted after initial procedures. Continuous variables were analyzed by Student's t test and categorical variables were analyzed by chi-square test. RESULTS: Three hundred and fifteen PVs were ablated in 85 patients underwent SPVA approach, 400 circular lesions surrounding ipsilateral PVs (including 790 PVs) were produced in the rest of 200 patients received CPVA approach. Electrical isolation was achieved in all of these PVs. Of these, PVPs were abolished in 89.8% (992/1105) of the ablated PVs, dissociated PV rhythms were documented in the rest 10.2 % (113/1105) of the treated PVs. Among the 113 dissociated PV rhythms, 28 met the criteria of PVT with mean CL of (155 +/- 43) ms (2 PVTs in 2 patients received SPVA, 26 PVTs in 18 patients underwent CPVA). PVT was more frequently documented in patients underwent CPVA approach [9.0% (18/200) vs 2.3% (2/85), P = 0.04]. During the 6-month follow-up, it was indicated that no significant difference existed in AF free rate between patients with PVT and those without PVT (P = 0.75). CONCLUSIONS: PVT dissociated from LA activations can be documented after PV isolation, especially in patients underwent CPVA approach. However, PVT does not affect the follow-up results.

[Pulmonary vein antrum isolation guided by 3-D mapping system in 100 patients with chronic atrial fibrillation].

OBJECTIVE: To investigate the efficacy and safety of circumferential pulmonary vein (PV) linear ablation (CPVA) guided by 3-D mapping system in patients with chronic atrial fibrillation (CAF). METHODS: From August 2004 to November 2005, 100 consecutive patients with CAF were admitted to undergo CPVA guided by CARTO system and EnSite NavX system, the main procedure end point is electrical isolation of PVs. Success was defined as atrial tachyarrhythmia free without any antiarrhythmia drugs for at least 3 months. Clinical and procedural variables were collected, ANOVA analysis was employed to identify the risk factors predicting recurrence, P < 0.05 was considered significant. RESULTS: After a mean of 9.7 +/- 5.7 months follow up, 70% (70/100) of patients freed from AF. ANOVA analysis identified isthmus ablation and poor left ventricular ejection fraction as the independent factors predicting success. Complications including pericardial tamponade (3 cases, 3%), stroke (1 case, 1%), asymptomatic pulmonary vein stenosis (2 cases, 2%). CONCLUSION: CPVA guided by 3-D mapping system can be performed in CAF patients with an acceptable efficacy, but safety need to be improved.

Characteristics in image integration system guiding catheter ablation of atrial fibrillation with a common ostium of inferior pulmonary veins.

BACKGROUND: Common ostium of the inferior pulmonary veins (PVs) is a kind of unusual variation in pulmonary venous drainage to the left atrium (LA), whose feature of anatomy, electrophysiology, and catheter ablation is rarely demonstrated, and the consecutive series of research for catheter ablation of atrial fibrillation (AF) in patients with that anomaly have not been reported. METHODS: A total of 1,226 patients with drug-refractory AF received magnetic resonance angiography (MRA) or multidetector computed tomography (MDCT) scan before ablation. Electrophysiological mapping was used to detect the focal triggers in paroxysmal AF. Basic catheter ablation strategy was circumferential PV isolation with "tricircle" under the guidance of image integration system: two circles surround two superior PVs, and the other surround the common trunk. RESULTS: LA and PVs reconstruction by image integration system showed a common pulmonary venous ostium of the right and left inferior PVs before ablation in 11 patients (0.9%). This anomaly could be classified into two types: type A without a short common trunk of inferior PVs and type B with a short common trunk. Fifty-seven percent paroxysmal AF was revealed focal triggers in the common ostium. The success rate of that strategy was 90%. CONCLUSION: Common ostium of inferior PVs could be classified into two types according to the presence of a short common trunk or not. The common ostium was usually an important triggering focus in paroxysmal AF. Catheter ablation strategy of circumferential PV isolation with "tricircle" under the guidance of image integration system would be a good choice.

Mechanisms of improvement of left ventricle remodeling by trans-planting two kinds of autologous bone marrow stem cells in pigs.

BACKGROUND: The necrosis of a large number of myocardial cells after acute myocardial infarction (AMI) results in a decrease of cardiac function and ventricle remodeling. Stem cell transplantation could improve cardiac function after AMI, but the involving mechanisms have not been completely understood. The present study aimed to investigate the effects of transplantation of autologous bone marrow mononuclear cells (BM-MNC) and mesenchymal stem cells (MSCs) via the coronary artery on the ventricle remodeling after AMI as well as the mechanisms of the effects of transplantation of different stem cells on ventricle remodeling. METHODS: A total of 36 male pigs were enrolled in this study, which were divided into 4 groups: control group, simple infarct model group, BM-MNC transplantation group, and MSCs transplantation group. At 90 minutes when a miniature porcine model with AMI was established, transplantation of autologous BM-MNC ((4.7 +/- 1.7) x 10(7)) and MSCs ((6.2 +/- 1.6) x 10(5)) was performed in the coronary artery via a catheter. Ultrasound, electron microscope, immunohistochemical examination and real time reverse transcriptase-polymerase chain reaction were used respectively to observe cardiac functions, counts of blood vessels of cardiac muscle, cardiac muscle nuclear factor (NF)-kappaB, myocardial cell apoptosis, and the expression of the mRNA of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in cardiac muscles. Multivariate Logistic regression was used to analyze the correlation factors of left ventricular end-diastolic diameter (EDD). RESULTS: The number of blood vessels in the infarct zone and around its border in the BM-MNC transplantation group was more than those in the infarct model group and MSCs group (P = 0.0001) and there was less myocardial cell apoptosis in the stem cell transplantation group than that in the infarct model group (all P < 0.01). The positive rate of NF-kappaB in the stem cell transplantation group was lower than that in the infarct model group (P = 0.001). The gene expression of VEGF in the infarct border zone of the BM-MNC group was higher than that in the MSCs group (P = 0.0001). The gene expression of bFGF in the infarct border zone in the MSCs transplantation group was higher than that in the infarct model group and the BM-MNC group (P = 0.0001). Left ventricular ejection fraction was inversely proportional to the apoptotic rate of myocardial cells and cardiac muscle NF-kappaB but positively correlated with the number of blood vessels and the expression of VEGF and bFGF in the infarct zone and infarct border zone. The Multivariate Logistic regression analysis on the factors influencing the left ventricular end-diastolic diameter after stem cell transplantation showed that the expression of VEGF mRNA in the cardiac muscles in the infarct zone, the number of apoptotic myocardial cells and the expression of NF-kappaB in the infarct border zone were independent factors for predicting the inhibitory effect on the dilation of left ventricular EDD after stem cell transplantation. CONCLUSIONS: Transplantation of autologous BM-MNC and MSCs in pigs can improve the condition of left ventricular remodeling and recover the cardiac functions after AMI. The improvement of cardiac functions is related to the increase of blood vessels, the increased expression of VEGF and bFGF, the reduction of myocardial cell apoptosis, and the decrease of NF-kappaB level in cardiac muscle tissues after stem cell transplantation.

Is circumferential pulmonary vein isolation preferable to stepwise segmental pulmonary vein isolation for patients with paroxysmal atrial fibrillation?

BACKGROUND: Stepwise segmental pulmonary vein isolation (SPVI) and circumferential pulmonary vein isolation (CPVI) have been developed to treat patients with atrial fibrillation (AF), but the preferable approach for paroxysmal AF (PAF) has not been established. METHODS AND RESULTS: One hundred and ten patients with symptomatic PAF were randomized into a stepwise SPVI group (n=55) or CPVI group (n=55). Systemic SPVI combined with left atrial linear ablation tailored by inducibility of AF was performed in the stepwise SPVI group. Circumferential linear ablation around the left and right-sided pulmonary veins (PVs) guided by 3-dimensional electroanatomic mapping was performed in the CPVI group. The endpoints of ablation are non-induciblity of AF in the stepwise SPVI group and continuity of circular lesions combined with PV isolation in the CPVI group. After the initial procedures, atrial tachyarrhythmis (ATa) recurred within the first 3 months in 23 of the 55 patients (41.8%) who underwent stepwise SPVI and in 20 of the 55 patients (36.4%) who had CPVI (p=0.69). Repeat procedures were performed in 7 patients from the stepwise SPVI group and 5 from the CPVI group (p=0.76). During the 3-9 months after the last procedure, 46 patients (83.6%) from the CPVI group and 43 (78.2%) from the stepwise SPVI group did not have symptomatic ATa while not taking anti-arrhythmic drugs (p=0.63). Severe subcutaneous hematoma or PV stenosis occurred in 3 patients. CONCLUSIONS: The efficacy of stepwise SPVI is comparable to that of CPVI for patients with PAF.